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AIM: To explore the effect of OCTN2 gene polymorphism on the expression and function of OCTN2, as well as the sensitivity of SW480 cells to oxaliplatin. METHODS: Four mutations of OCTN2 (F17L, E317K, S467C and P478L) transfected cell lines were constructed. Real-time RT-PCR and Western blot were used to detect the levels of OCTN2 mRNA and protein. The content of oxaliplatin was detected by HPLC. MTS assay was used to detect cell viability. RESULTS: The expression level of all mutant OCTN2 mRNA and protein was not significantly different from that of wild-type OCTN2. Oxaliplatin uptake experiments showed that there was no significant difference in V
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AIM:To probe whether CpG oligodeoxyribonucleotides 7909 (CpG ODN7909) combined with Toll like receptor (TLR)9 affected the chemotherapeutic sensitivity of doctaxel (DOC) in human lung cancer A549 and H520 cell lines.METHODS:Sequences of TLR9 siRNAs were designed.A549 and H520 cells were transfected with TLR9 siR-NA by lipofectamine.The expression of TLR9 was detected by Western blot .The cell activity was measured by CCK-8 as-say.The experiments were divided into blank control group , control siRNA group and TLR9 siRNA interference group.The cell cycle distribution and cell apoptosis were analyzed by flow cytometry .The expression of P38 and Bax was determined by Western blot .The cells in each group were exposed to CpG ODN 7909 and/or DOC.RESULTS: In A549 cells and H520 cells, CpG ODN7909 alone had no obvious effect on the cell activity , G2/M phase arrest and apoptosis , but in-creased the protein expression of P 38 and Bax ( P<0.01) .In addition, there was no significant changes of the above inde-xes in CpG ODN7909 treated-TLR9 siRNA group was observed .DOC alone significantly inhibited the cell activity , higher the G2/M phase fractions, apoptotic rates and Bax expression (P<0.01), but didn’t affect the expression of P38 in all 3 groups.Compared with the cells treated with DOC alone , the cells treated with CpG ODN7909 combined with DOC exhibi-ted lower cell activity, higher G2/M phase fractions, apoptosis rates and more Bax expression (P<0.01), showed no sig-nificant change of P38 expression.In addition, there was no significant change of the above indexes in CpG ODN 7909 com-bined with DOC treated-TLR9 siRNA group was observed .CONCLUSION:CpG ODN7909 may enhance the chemothera-peutic sensitivity of DOC in human lung cancer cells by combining with TLR 9.The mechanism might be related to enhan-cing the inhibitory effect and apoptosis of DOC on the cell activity in vitro, arresting the cells at G 2/M phase of the cells .
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AIM:To investigate whether Notch1 changes stemness and chemotherapeutic sensitivity in human glioma U251 cells.METHODS: The lentiviral vectors, which expressed Notch1-shRNA or Notch1 intracellular domain ( NICD) , were transfected into U251 cells .Western blot and immunofluorescence staining were applied to monitor the va-lidity of the cells, down-regulation of Notch1 expression or over-expression of NICD.The proportion of CD133 +cells was analyzed by flow cytometry.The expression of nestin and GFAP was identified by immunofluorescence staining.The forma-tion rate of tumor cell spheres and the implanted tumor growth in SCID mice were observed.MTT assay was performed to e-valuate the chemotherapeutic sensitivity to VM-26 and BCNU of the cells with different treatments.RESULTS:Stemness was significantly enhanced in the cells over-expressing NICD.For example, the proportion of CD133 +cells was increased, the expression of nestin was up-regulated, the expression of GFAP was down-regulated, and the formation rate of tumor cell spheres and implanted tumor growth were increased.The chemotherapeutic sensitivity to VM-26 and BCNU of the cells was decreased.In the cells with Notch1 gene down-regulation by RNAi, the stemness was inhibited and chemotherapeutic sensi-tivity was increased.CONCLUSION:Notch1, which leads to the change of stemness and chemotherapeutic sensitivity in human glioma U251 cells, is likely to be a potential molecular target for treatment of glioma.
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AIM:To investigate the role of Herceptin in the apoptosis and drug sensitivity of endometrial canc -er Ishikawa cells .METHODS: The IC50 values of Herceptin , adriamycin ( ADR ) , cisplatin ( DDP ) and paclitaxel ( PTX) for Ishikawa cells were detected by MTT method .Ishikawa cells were treated with single drug and combined chemo-therapy for 24 h, the cell cycle and the apoptosis ratio were determined by flow cytometry .RESULTS:The IC50 values of Herceptin, ADR, DDP and PTX were 57.12 mg/L, 0.572μmol/L, 67.4μmol/L and 719.5 nmol/L, respectively.Her-ceptin significantly enhanced the cytotoxicity of the chemotherapeutic drugs , and increased apoptosis ratio statistically . CONCLUSION:Herceptin enhances the apoptosis-inducing ability of the chemotherapeutic drugs and improves the che-motherapeutic sensitivity in Ishikawa cells .
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Objective:To investigate the correlation of plasma D-dimer levels with the response to first-line chemotherapy and the prognosis of patients with serous ovarian cancer (SOC). Methods:The preoperative plasmic D-dimer levels of 143 patients with prima-ry SOC were retrospectively evaluated. The patients were admitted to Tianjin Medical University Cancer Institute and Hospital between January 2008 and May 2010. The patients were divided into two groups on the basis of plasmic D-dimer levels. Group A consisted of 100 patients with a normal plasmic D-dimer level of≤0.3 mg/L. Group B included 43 patients with an increased plasmic D-dimer level of>0.3 mg/L. The correlations of the different plasmic D-dimer levels with clinicopathological features, therapeutic effects, and surviv-al outcomes were further analyzed. Results:The plasmic D-dimer levels were positively correlated with the staging of the Federation of International Gynecology and Obstetrics, residual tumor size, presence of malignant ascites, preoperative serum CA125 level, and neo-adjuvant chemotherapy. Group B exhibited a significantly lower (P<0.001) complete response (CR) rate of 34.88%(15/43) than group A, which yielded a CR rate of 73.00%(73/100). The progression-free survival and overall survival rates of group B were significantly lower than those of group A (25.58%vs. 50.00%and 32.56%vs. 65.00%;P<0.05). Multivariate analysis revealed that the plasmic D-di-mer level is an independent prognostic factor associated with unfavorable prognosis. Conclusion:Increased preoperative plasmic D-di-mer levels may be a potential biomarker of weak responses to first-line chemotherapy and poor clinical outcomes in patients with SOC.
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OBJECTIVE: To explore the effects of sodium nitrite pretreatment on the chemotherapeutic sensitivity to adriamycin (ADM) in MG63 cells.
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Objective To detect the expression of BAG-1 and p53 in postoperative patients paraffin section of non-snmll cell lung cancer (NSCLC), and retrospectively analyze the relationships of clinical feature and platinum-based chemotherapeutic sensitivity in NSCLC, to further investigate the resistance mechanism of platinum. Methods The expression of BAG-1 and p53 was measured by immunohistochemistry in paraffin sections of 125 NSCLC postoperative patients. The existence and prognosis were analyzed after follow-up. Results BAG-1 and p53 showed high expression in NSCLC (positive rate 64.8 %, 46.7 %); BAG-1 positive or p53 negative expression had survival advantage(P<0.05); The group of BAG-1, p53 negative expression could benefit from platinum-based chemotherapy (P<0.05). Conclusion Expressions of BAG-1, p53 correlates with platinum-based chemotherapeutic sensitivity, and maybe become new types of clinical prognostic indicators and contribute to individualized treatment options for NSCLC.
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The expression of X-linked inhibitor of apoptosis protein (XIAP) gene and its effect on chemotherapeutic sensitivity in bladder carcinoma was explored. By using immunohistochemistry,the expression of XIAP was detected in 47 bladder carcinomas and 5 normal bladder tissues. The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418. Cellular XIAP mRNA level was detected by RT-PCR. Low-dose mitocycin C was administered to induce the apoptosis of T24 cells. The in vitro growth of bladder carcinoma cells was analyzed by MTT colorimetry, and the apoptosis rate was assayed by TUNEL methods. It was found XIAP was moderately expressed in bladder carcinomas with the the positive rate being 78.73% (37/47), but the positive rate was not correlated with carcinoma stages and grades (P＜0.05). XIAP mRNA level in transfected T24 cells was significantly increased by 3.8 times as compared with that in the cells not transfected with XIAP. After treatment with low-dose mitomycin C (0.005 and 0.05 mg/mL), the growth rate in XIAP no-transfected control group was increased by (11.60±0.25)% and (16.51±0.87)% (P＜0.05), and the apoptosis rate was decreased by (10.1±0.2)% and (11.9±0.2%) (P＜0.05) respectively as compared with XIAP transfected group. It was concluded that XIAP was expressed in most of bladder carcimoma samples. Overexpression of XIAP in T24 could significantly reduce the MMC-induced apoptosis of bladder carcinoma, suggesting its effect on the chemotherapeutic sensitivity of T24 cells.
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Objective To explore the investigate of X-chromosome-linked inhibitor of apoptosis pro- tein(XIAP)and its effect on chemotherapeutic sensitivity in bladder carcinoma.Methods Using immu- nohistochemistry methods,the expression of XIAP was evaluated in 47 bladder carcinomas and 6 normal bladder tissues.The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418.Cellular XIAP mRNA level was detected by RT-PCR.The apoptosis of T24 cells was induced by low-dose of mitocycin C(0.005 mg/ml and 0.05 mg/ml,respectively).The in vitro cellular growth activities were assayed by MTT color imetry;and the apoptosis rate was assayed by TUNEL methods. Results The expression rate of XIAP was 78.7%(37/47)in bladder carcinoma samples,with no corre- lation with carcinoma stages and grades(P＞0.05).XIAP mRNA level in transfected T24 ceils was signifi- cantly increased by 3.8 times.Treated with 0.005 mg/ml and 0.05 mg/ml of mitomycin C,the growth rates of XIAP transfected T24 cells were increased [(11.60?0.25)% and(16.51?0.87)% ,respectively,P＜0.05];and the apoptosis rates were decreased [(10.1?0.2)% and( 11.9?0.2)% ,respectively,P＜0.05]compared with those in control cells.Conclusions XIAP is highly expressed in humun bladder car- cinoma samples.Overexpression of XIAP in T24 cells results in decrease in bladder carcinoma cell apoptosis induced by MMC,which may decrease the chemotherapeutic sensitivity of T24 cells.
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Objective:To study the feasibility and reliability of using phosphorylated H2AX(?H2AX)as a predictor for sensitivity of hepatic carcinoma cell HepG2.215 to chemotherapy agents: etoposide, doxorubicin, mitomycin, and cisplatin. Methods: HepG2.215 cells were exposed to etoposide, doxorubicin, mitomycin or cisplatin of 1, 2, 4 and 20 concentration index (CI). Untreated HepG2.215 cells were taken as control. The proportion of HepG2.215 cells expressing ?H2AX was measured by flow cytometry, the number of ?H2AX foci in HepG2.215 cells was measured by immunocytochemistry, and cell proliferation was measured by MTT. The correlation between the number of ?H2AX foci and the percentage of HepG2.215 cells expressing ?H2AX in HepG2.215 cells was analyzed; the correlation of CI with the percentage of HepG2.215 cells expressing ?H2AX or ?H2AX foci and inhibitory rate of cell proliferation was analyzed; and the correlation of inhibitory rate of cell praliferation with the percentage of HepG2.215 cells expressing ?H2AX or ?H2AX foci was also analyzed. Results: There was a positive correlation between the number of ?H2AX foci and the percentage of HepG2.215 cells expressing ?H2AX in HepG2.215 cells after treatment with the above 4 agents (all P